Development of simple and efficient in planta transformation method for rice (Oryza sativa L.) using Agrobacterium tumefaciens

Supartana, Putu; Shimizu, Tsutomu; Shioiri, Hidenari; Nogawa, Masahiro; Nozue, Masayuki; Kojima, Mineo

doi:10.1263/jbb.100.391

Cited by 108 publications

(57 citation statements)

References 14 publications

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“…K-134, transgenics were developed by a tissue culture independent Agrobacterium-mediated in planta transformation strategy. The in planta transformation protocols have been developed for transformation in many crop species, Arabidopsis thaliana (Feldmann and Marks 1987), soybean (Trick and Finer 1997), sunflower (Rao and Rohini 1999), safflower (Rohini and Rao 2000), rice (Supartana et al 2005), maize (Chumakov et al 2006) and cotton (Keshamma et al 2008). Earlier an in planta transformation protocol was developed in groundnut by Rohini and Rao (2000) and proved its reproducibility by Keshamma et al (2008).…”

Section: Discussionmentioning

confidence: 99%

Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Keshavareddy

Rohini

Ramu

et al. 2013

Physiol Mol Biol Plants

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Large number of primary transgenic events were generated in groundnut by an Agrobacterium mediated, in planta transformation method to assess the efficacy of cry1AcF against the Spodoptera litura. generation demonstrated homozygous nature. This clearly proved that though there is considerable improvement in average mean % larval mortality in T 2 generation, the cry1AcF gene was effective against S. litura only to some extent.

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Section: Discussionmentioning

confidence: 99%

Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Keshavareddy

Rohini

Ramu

et al. 2013

Physiol Mol Biol Plants

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“…These methods have been successfully used for transformation of important genes to many plants whose tissue culture system has already been established (Hiei et al 1994;Ishida et al 1996;Gelvin 2003;Carneiro and Carneiro 2013;Martins et al 2015;Jan et al 2016). Some advantages of transformation via A. tumefaciens have been reported such as higher transformation efficiency with a single copy gene and can be carried out with simple laboratory equipment and low-cost technique (Supartana et al 2005). Transformants with a single copy gene are preferable as it is a easest analysis process and the segregation usually follows Mendelian inheritance pattern (Rahmawati 2006).…”

Section: Introductionmentioning

confidence: 99%

“…Supartana et al (2005) obtained the levels of efficiency of in-planta transformation of 40% (PCR) and 34% (GUS histochemical test) by inoculation of embryonic apical meristem rice plants. In-planta transformation using filament pistil (female flowers) of maize with plasmid pTd33 containing gus and nptII genes showed that 6.8% sprouts from the seeds were resistant to kanamycin and 60.3% of the resistant gene nptII were positive (Chumakov et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium tumefaciens-MEDIATED IN-PLANTA TRANSFORMATION OF INDONESIAN MAIZE USING pIG121Hm-Cs PLASMID CONTAINING nptII AND hpt GENES

Listanto¹,

Riyanti²,

Sustiprijatno³

2017

Indones. J. Agric. Sci.

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Maize (Zea mays L.) productivity in Indonesia is challenged to be increased using genetic engineering. Recent advances in Agrobacterium tumefaciens-mediated in-planta transformation makes it possible to transform maize with low cost, and simple method. This study aimed to confirm pIG121Hm-Cs plasmid in A. tumefaciens, and to estimate the efficiency level of A. tumefaciens-mediated in-planta transformation of Indonesian maize by using pIG121Hm-Cs plasmid containing nptII and hpt genes. A series of studies were conducted including confirmation of gene construct of pIG121Hm-Cs plasmid in A. tumefaciens, transformation of four maize lines through A. tumefaciens-mediated in-planta technique, acclimatization of transformant plants and molecular analysis of selected plants using polymerase chain reaction (PCR). The pIG121Hm-Cs plasmid was confirmed via PCR analysis using specific primers of nptII and hpt genes and resulted 700 bp and 500 bp for fragments of nptII and hpt, respectively. After selection, acclimatization and molecular analysis steps, the efficiency levels of transformation of four maize lines were low, ranging from 3.8% to 12.8%. The level of transformation efficiency of ST-27 line was the highest accounting for 12.8% of 45 planted embryos on selection medium based on PCR analysis using specific primer for nptII gene. Overall, A. tumefaciensmediated in planta transformation on maize floral pistil in this study proved to be successful and rapid. Therefore, this enhanced transformation method will be beneficial for Indonesian maize genetic engineering.

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“…Today, the method is applied to many plants, including Arabidposis, rice, maize and wheat (Feldmann and David, 1987;Supartana et al, 2005Supartana et al, , 2006. This method eliminates tissue culture process and only requires soaking mature seeds in Agrobacterium tumefaciens.…”

Section: Introductionmentioning

confidence: 99%

Development of a Simple and Efficient Method for Agrobacterium-Mediated Transformation in Sorghum

Li¹,

Wang²,

Zhan³

et al. 2015

IJAB

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Sorghum is considered a recalcitrant crop based on genetic transformation. The main technical problem is callus browning in culture tissue and genetic transformation for sorghum. In this study, we developed a simple and effective method for sorghum transformation excluding tissue culture process. Sorghum mature embryos were pierced by a needle and then soaked in an Agrobacterium inoculum with acetosyringone. The best-performing concentration of acetosyringone was 100 μM in inoculation with pierced seeds. When these inoculated seeds grew to three leaves, a 1‰ hygromycin solution was used to brush the second and third leaf of inoculated seedlings to identify transformants. The identification results showed that the highest efficiency of transformation was 4% in the study. Molecular analysis for T0 and T1 plants identified that target gene was integrated into sorghum genomic DNA. And target gene can normally express in transformants' leaves. Genetic analysis of T1 transformants showed that most of T1 transformants fitted to 3:1 Mendelian inheritance. All these results showed that the presented method is an easy and successful method for sorghum transformation. In the future, the method will be used for broad functional genomics studies and for biotechnological application of sorghum.

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Development of simple and efficient in planta transformation method for rice (Oryza sativa L.) using Agrobacterium tumefaciens

Cited by 108 publications

References 14 publications

Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Agrobacterium tumefaciens-MEDIATED IN-PLANTA TRANSFORMATION OF INDONESIAN MAIZE USING pIG121Hm-Cs PLASMID CONTAINING nptII AND hpt GENES

Development of a Simple and Efficient Method for Agrobacterium-Mediated Transformation in Sorghum

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